MOST CLOSELY HLA MATCHED ALLOGENEIC VIRUS SPECIFIC CYTOTOXIC T-LYMPHOCYTES (CTL)

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 摘要 当患者接受来自捐赠者的造血干细胞移植(HSCT)时，如果捐赠者是治疗血癌或其他血液疾病的最佳配型，他们就有感染的风险，直到他们能够从捐赠者那里培养出新的免疫系统。在此期间，他们可能会患上严重的病毒感染，爱泼斯坦-巴尔病毒(EBV)、巨细胞病毒和腺病毒是最常见的三种可能导致问题的病毒。研究人员此前已经证明，从移植捐赠者那里培养出称为病毒特异性CTL的特殊T细胞是可能的，当它们被回馈给患者时，可以预防和治疗这些病毒感染。然而，培养这些细胞需要2-3个月的时间，所以为每个感染这些病毒的患者培养来自移植捐赠者的病毒特异性CTL是不现实的选择。 在这项研究中，研究人员将研究是否有另一种方法是让来自正常捐赠者的病毒特异性CTL库冷冻保存，然后在移植后提供给患有此类病毒感染的受试者进行治疗。病毒特异性CTL株将从正常捐赠者身上培养并冷冻。为了建立CTL系，我们首先用一种专门生产的腺病毒(一种载体)感染称为单核细胞的血细胞，这种病毒也携带部分巨细胞病毒(CMV)基因。这是一种被禁用的病毒，一旦感染发生，就不能自我复制，因此不能传播。这些被感染的单核细胞然后刺激T细胞对腺病毒和CMV做出反应，并杀死感染这些病毒的细胞。然后，我们对T细胞进行第二次刺激，使用也感染了EBV的细胞(我们将在实验室通过感染EBV从捐赠者的血液中提取EBV)，以便细胞现在识别三种病毒：CMV、EBV和腺病毒。一旦我们制造了足够数量的T细胞，我们将对它们进行测试，以确保它们杀死感染这些病毒的细胞并将其冷冻。 如果移植患者感染了这些病毒中的一种，并且尽管进行了标准治疗，但感染仍然存在，他或她将有资格接受适当匹配的CTL株。 主要目标是确定这一策略是否可行和安全。一种风险是，因为我们制造的T细胞与受者的基因(HLA)不完全匹配，它们可能会攻击受试者，导致一种称为移植物抗宿主病(GVHD)的疾病。我们将密切关注这一并发症。 第二个目标是确定T细胞输注对病毒感染的影响，并看看接受者是否能对这些病毒保持免疫力。我们还将观察T细胞存活多长时间。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. ABSTRACT When patients undergo a hemopoietic stem cell transplant (HSCT) from a donor who is the best possible match to treat blood cancers or other blood diseases they have a risk of infection until they can grow a new immune system from the donor. During this period they may develop serious viral infections with Epstein Barr virus (EBV), cytomegalovirus and adenovirus being three of the most common viruses that can cause problems. Investigators have previously shown that it is possible to grow up special T cells called virus-specific CTLs from the transplant donor that can prevent and treat these viral infections when they are given back to the patient. However it takes 2-3 months to grow these cells so it is not a practical option to grow virus-specific CTLs from the transplant donor for every patient that gets an infection with one of these viruses. In this study, investigators will see if an alternative approach is to make banks of virus-specific CTLs from normal donors that could be stored frozen and then made available to treat subjects post transplant if they developed one of these viral infections. Virus-specific CTL lines will be grown from normal donors and frozen. To make the CTL lines we first infect blood cells called monocytes with a specially produced adenovirus (a vector) that also carries part of the Cytomegalovirus (CMV) gene. This is a disabled virus that cannot reproduce itself once infection has occurred so it cannot spread. These infected monocytes then stimulate the T cells to respond to adenoviruses and CMV, and kill the cells infected with these viruses. We then give a second stimulation to the T cells, using cells which are also infected with EBV (which we will make from donor blood by infecting them with EBV in the laboratory) so that the cells now recognize three viruses CMV, EBV and Adenovirus. Once we have made sufficient numbers of T cells we will test them to make sure they kill cells infected with these viruses and freeze them. If a transplant patient gets an infection with one of these viruses, and the infection persists despite standard therapy, he or she would be eligible to receive a suitably matched CTL line. The primary objective is to determine if this strategy is feasible and safe. One risk is that because the T cells we make are not a complete genetic (HLA) match for the recipient, they might attack the subject and cause a condition called graft versus host disease (GVHD). We will closely monitor for this complication. Secondary objectives are to determine the effects of the T cell infusion on the virus infection and to see if the recipients can stay immune to these viruses. We will also see how long the T cells survive.